The HSCR samples were excised from normal and aganglionic segment and then fixed in polyoxymethylene (4%) for at least 48 h. Fixed samples were washed and dehydrated by gradient ethanol, embedded in paraffin and sliced. To analyse the expression location of target DEGs in HSCR samples, IHC for MFAP5 (Abcam,
ab203828, Rabbit monoclonal immunoglobulin G (IgG), 1:5000 dilution), THBS4 (Abcam,
ab263898, Rabbit monoclonal IgG, 1:600 dilution), POSTN (Abcam,
ab215199, Rabbit monoclonal IgG, 1:1000 dilution), ANXA1 (Abcam,
ab214486, Rabbit monocle IgG, 1:6000 dilution), HSP70 (Abcam,
ab181606, Rabbit monoclonal IgG, 1:1000 dilution), HSP40 (Cell Signaling Technology, 4871T, Rabbit monoclonal IgG, 1:50 dilution), NR2F1 (Abcam,
ab181137, Rabbit monoclonal IgG, 1:100 dilution) and ZEB2 (Santa Cruz Biotechnology,
sc‐271984, mouse monoclonal IgG, 1:50 dilution) was performed on 4‐μm‐thick paraffin‐embedded formalin‐fixed tissue. The
GTVisionTM III Detection System/Mo&Rb kit (GENE, GK500710) was used for IHC detection according to the manufacturer's protocol. Slides were imaged using a Leica DM750 ideal microscope with the Leica
ICC50 HD camera.
He S., Wang J., Huang Y., Kong F., Yang R., Zhan Y., Li Z., Ye C., Meng L., Ren Y., Zhou Y., Chen G., Shen Z., Sun S., Zheng S, & Dong R. (2023). Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing. Clinical and Translational Medicine, 13(2), e1193.
